Bio-Rad Experion Protein Analysis Kits - Manual (Page 23)

2.3 Data Analysis
Evaluate the run and the analysis of the data by Experion software.
1. Evaluate the virtual gel to ensure that all lanes (samples) are visible. Check that the
lower marker is present in each sample (indicated by pink triangles) and that it is
aligned across all lanes (Figure 2.2).
2. Select Graph > Scale to Global to adjust the fluorescence intensity scale so that the
virtual gel most closely resembles a gel-based separation (Figure 2.2). All lanes are
scaled to the highest peak found across the entire chip, which enables comparison of
the concentration differences among the samples in the virtual gel.
Fig. 2.2. Virtual gel generated by Test 2. L, RNA ladder; lanes 1, 4, 7, and 10, 25 ng/µl total RNA;
lanes 2, 5, 8, and 11, 100 ng/µl total RNA; lanes 3, 6, 9, and 12, 500 ng/µl total RNA. In this example, the
virtual gel was “scaled to global” to emphasize the differences in sample concentrations.
3. Evaluate the separation of the RNA ladder. This is a critical step in data analysis
because much of the automated data analysis performed by Experion software is
based on the successful separation of the RNA ladder. To display the ladder
electropherogram, click the ladder well in the project tree, or click on the lane labeled L
in the virtual gel. The electropherogram should resemble the one shown in Figure 1.7.
4. Examine the separation of one of the total RNA samples. Click on a sample name in
the project tree or on a lane in the virtual gel. The electropherogram appears and
should have the features described for Figure 1.8 (a lower marker peak at ~21 sec that
is at least 5 fluorescence units above the baseline and distinguishable from the sample
peak, a flat baseline, and well-resolved 18S and 28S rRNA peaks).
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